LOCUS       sequence-218373-        2626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Downstream homology arm, amyE encodes _-amylase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2626)
  AUTHORS   Wicke N, Radford D, Verrone V, Wipat A, French CE
  TITLE     BacilloFlex: A modular DNA assembly toolkit for Bacillus subtilis 
            synthetic biology
  JOURNAL   bioRxiv 185108
REFERENCE   2  (bases 1 to 2626)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            185108"
COMMENT     Sequence Label: pTU1-D-amyED
FEATURES             Location/Qualifiers
     source          1..2626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(574..591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(745..1333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(825..844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1504..2295,2296..2364))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2127..2146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2365..2469)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2537..2555
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2559..2602)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2605..2624
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
ORIGIN
        1 cgtctcagaa gttaggctgg gcggtgatag cttctcgttc aggcagtacg cctcttttct
       61 tttccagacc tgagggaggc ggaaatggtg tgaggttccc ggggaaaagc caaataggcg
      121 atcgcgggag tgctttattt gaagatcagg ctatcactgc ggtcaataga tttcacaatg
      181 tgatggctgg acagcctgag gaactctcga acccgaatgg aaacaaccag atatttatga
      241 atcagcgcgg ctcacatggc gttgtgctgg caaatgcagg ttcatcctct gtctctatca
      301 atacggcaac aaaattgcct gatggcaggt atgacaataa agctggagcg ggttcatttc
      361 aagtgaacga tggtaaactg acaggcacga tcaatgccag gtctgtagct gtgctttatc
      421 ctgatgatat tgcaaaagcg cctcatgttt tccttgagaa ttacaaaaca ggtgtaacac
      481 attctttcaa tgatcaactg acgattacct tgcgtgcaga tgcgaataca acaaaagccg
      541 tttatcaaat cttagagaga cgctgcaggc ttcctcgctc actgactcgc tgcgctcggt
      601 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      661 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      721 taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg agcatcacaa
      781 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      841 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      901 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      961 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1021 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1081 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1141 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1201 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1261 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1321 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1381 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1441 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1501 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1561 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1621 ccccagtgct gcaatgatac cgcgagagcc acgctcaccg gctccagatt tatcagcaat
     1681 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1741 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1801 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1861 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1921 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1981 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2041 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2101 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2161 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2221 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2281 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2341 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2401 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2461 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     2521 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
     2581 agctttcgct aaggatgatt tctggaattc gcggccgctt ctagaa
//
